Exploring Toxoplasma gondii Allelic and Haplotypic Diversity in Wildlife from Panama Using the alt SAG2, GRA6, and Apico Markers

Background: Panama is recognized as a global biodiversity hotspot; however, little is known about the genetic diversity patterns of Toxoplasma gondii in wildlife. This exploratory study evaluated the allelic and haplotypic diversity of the alt SAG2, GRA6, and Apico markers using wildlife tissue samples; additionally, it compared the genealogical relationships between the identified variants and a defined panel of reference sequences. Methods: Amplicons from each tissue sample and locus collected from 13 wild animals were analyzed using Sanger sequencing. The resulting sequences were then examined using NeighborNet networks and unrooted Neighbor-Joining trees. Results: A total of 162 sequences were generated for each locus. The alt SAG2 variants clustered with reference alleles associated with T. gondii types II and III, whereas GRA6 variants clustered with reference alleles associated with types I, II, and III. The Apico fragment did not separate the type I, II, and III reference sequences. Multiple sequence variants were recovered from several sets of clones derived from the same tissue. Conclusions: The genetic variation detected in Panamanian wildlife comprises a heterogeneous set of T. gondii sequence variants associated with archetypal clonal variants and with sequence variants described in geographically distant regions. These findings suggest that the allelic variation present in the analyzed wildlife is not restricted to the regional context but is part of a broader pattern of T. gondii diversity in the Neotropics.